Details of experiments between Q86SQ6 and Q86UL8
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ADGRA1
Protein A fragment: 551-560
Protein A site: PBM
Protein A sequence: TGPWKNETTV
Protein A construct: biotin-ado-ado-TGPWKNETTV
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.62
Normalized holdup BI: 0.69
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.17
Experiment series 2
Protein A protein: ADGRA1
Protein A fragment: 551-560
Protein A site: PBM
Protein A sequence: TGPWKNETTV
Protein A construct: biotin-ado-ado-TGPWKNETTV
Protein B protein: MAGI2
Protein B fragment: 772-865
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.35
Normalized holdup BI: 0.47
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.74
Experiment series 3
Protein A protein: ADGRA1
Protein A fragment: 551-560
Protein A site: PBM
Protein A sequence: TGPWKNETTV
Protein A construct: biotin-ado-ado-TGPWKNETTV
Protein B protein: MAGI2
Protein B fragment: 915-1017
Protein B site: PDZ_5
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.15
Normalized holdup BI: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.02
Experiment series 4
Protein A protein: ADGRA1
Protein A fragment: 551-560
Protein A site: PBM
Protein A sequence: TGPWKNETTV
Protein A construct: biotin-ado-ado-TGPWKNETTV
Protein B protein: MAGI2
Protein B fragment: 600-682
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.12
Normalized holdup BI: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.94
Experiment series 5
Protein A protein: ADGRA1
Protein A fragment: 551-560
Protein A site: PBM
Protein A sequence: TGPWKNETTV
Protein A construct: biotin-ado-ado-TGPWKNETTV
Protein B protein: MAGI2
Protein B fragment: 1141-1229
Protein B site: PDZ_6
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
Normalized holdup BI: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.71